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Development of a simple HPLC–MS/MS method to simultaneously determine teriflunomide and its metabolite in human plasma and urine: Application to clinical pharmacokinetic study of teriflunomide sodium and leflunomide
Author(s) -
Yao Xueting,
Liu Yang,
Song Ling,
Jiang Ji,
Xiao Fei,
Liu Dongyang,
Hu Pei
Publication year - 2019
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.4420
Subject(s) - teriflunomide , chemistry , chromatography , protein precipitation , metabolite , urine , active metabolite , leflunomide , pharmacokinetics , electrospray ionization , high performance liquid chromatography , tandem mass spectrometry , mass spectrometry , pharmacology , multiple sclerosis , fingolimod , medicine , psychology , biochemistry , methotrexate , psychiatry , biology , immunology
A simple high‐performance liquid chromatography coupled with tandem mass spectrometry method was developed and fully validated to simultaneously determine teriflunomide (TER) and its metabolite 4‐trifluoro‐methylaniline oxanilic acid (4‐TMOA) in human plasma and urine. Merely 50 μL plasma and 20 μL urine were employed in sample preparation using protein precipitation and direct dilution method, respectively. An Agilent Zorbax eclipse plus C 18 column was selected to achieve rapid separation for TER and 4‐TMOA within 3 min. Electrospray ionization under multiple reaction monitoring was used to monitor the ion transitions for TER ( m/z 269.0 → 159.9), 4‐TMOA ( m/z 231.9 → 160.0), internal standard teriflunomide‐d4 ( m/z 273.0 → 164.0) and 2‐amino‐4‐trifluoromethyl benzoic acid ( m/z 203.8 → 120.1), operating in the negative ion mode. This method proved to have better accuracy and precision over concentration range of 10–5000 ng/mL in plasma as well as 10–10,000 ng/mL in urine. After a full validation, this method was successfully applied in a pharmacokinetic study of teriflunomide sodium and leflunomide in Chinese healthy volunteers.